Impact Of Host Plant Xylem Fluid On Xylella Fastidiosa Multiplication ...

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OBJECTIVES The overall objectives of our research are to: 1. Quantify GWSS and natural enemy dispersal patterns in a complex landscape and 2. Determine which factors influence their dispersal. To accomplish these objectives we must first develop a mark-capture protein marking technique and quantify the protein marking retention intervals for the targeted insects. Field application of better mark-capture techniques will enhance our understanding of the area-wide dispersal patterns of GWSS and its natural enemies. RESULTS Direct Contact Marking Method Dozens of nylon-meshed sleeve cages (66 X 70-cm, 18-cm dia.) were placed on randomly selected citrus branches located at the Agricultural Operations Research Station in Riverside, CA. Adult GWSS and H. convergens were then introduced into each cage and sprayed with a 5.0% solution of non-fat dry milk (NFDM) or chicken egg whites (All Whites). A single cage from each marking treatment was randomly selected on 12 different sampling dates for up to 35 days after marking. All of the surviving GWSS and H. convergens in the randomly selected cages were assayed by an anti-NFDM or an anti-egg white ELISA to detect the presence of each respective protein mark. Residual Contact Marking Method Randomly selected citrus branches located at the Agricultural Operations Research Station in Riverside, CA were sprayed with a 5.0% solution of NFDM or chicken egg whites. The branches were allowed to dry for several hours, and then nylonmeshed sleeve cages were placed on the branches. Adult GWSS and H. convergens were then introduced into each cage. The sampling scheme was the same as the one described above. All of the surviving GWSSs and H. convergens in the randomly selected cages were assayed by an anti-NFDM or an anti-egg white ELISA to detect for the presence of each respective protein marker. The ELISA results for the protein marked GWSS are given in Table 1. Data indicate that both marking procedures, regardless of the type of protein marker used, were retained well on GWSS. As expected, the topical marking procedure yielded higher ELISA values and had longer retention than the residual contact marking method. Generally, the markers were retained on 100% of the GWSS for ≈ 2 and 3 weeks by the residual and topical marking procedures, respectively. The ELISA results for the protein-marked H. convergens are given in Table 2. H. convergens ELISA reactions were very similar to the reactions yielded by GWSS. CONCLUSIONS In the first phase of our research described here, we showed that protein markers can be retained on insects several weeks after marking in the field. This marking technique provides the necessary tool to distinguish GWSS and its natural enemies so that studies of dispersal, migration, longevity, and density can be conducted. Additionally, different protein markers can be used to identify insects released at different times, in different areas, or in different crops. Next, we will use this technique to investigate the landscape-level movement of GWSS (nymphs and adults) and its natural enemies. We propose to use the mark-capture system to simultaneously quantify the intercrop dispersal of GWSS and its natural enemies. Specifically, we will spray large areas (e.g., field plots, whole trees, bushes, etc.) with inexpensive proteins using conventional spray equipment. In turn, insects that are hit by the protein solutions or that eat or walk on plant material containing protein residues will obtain enough protein to be detected by protein-specific ELISAs. Because the two marking ELISAs (chicken egg whites and NFDM) do not cross-react, we can apply the materials to two different host plants in close proximity to one another. Then, insects can be collected using temporal and spatial sampling schemes and analyzed for the presence of each respective protein marker to determine not only the insect’s point of origin but the timing and extent to which portions of the population move among different plant species. FUNDING AGENCIES Funding for this project was provided by the University of California Pierce’s Disease Grant Program and the USDA Agricultural Research Service. - 257 -
Table 1. The mean (±SD) ELISA readings and the percentages of protein-marked GWSS scoring positive for the presence of chicken egg white or non fat dry milk for up to 35 days after marking. GWSS were scored positive for the presence of each marker if the ELISA value exceeded the mean negative control value by 3 standard deviations. Egg White Marker Non Fat Dry Milk Marker Days Mean Mean Application Method After Marking Number Assayed ELISA Reading Percent Positive Number Assayed ELISA Reading Percent Positive Residual Contact 1 31 0.49 (0.3) 100.0 8 0.38 (0.2) 100.0 3 7 0.46 (0.4) 100.0 10 0.38 (0.2) 100.0 5 19 0.94 (0.4) 100.0 4 0.43 (0.1) 100.0 8 15 0.71 (0.3) 100.0 5 0.20 (0.1) 100.0 12 26 0.57 (0.4) 88.5 36 0.36 (0.2) 100.0 13 7 0.52 (0.3) 100.0 5 0.28 (0.3) 100.0 15 26 0.31 (0.2) 100.0 6 0.27 (0.3) 83.3 17 13 0.40 (0.2) 100.0 15 0.11 (0.1) 66.7 19 13 0.17 (0.2) 76.9 5 0.11 (0.1) 40.0 21 3 0.10 (0.1) 66.7 6 0.08 (0) 66.7 34 0 --- --- 3 0.06 (0) 33.3 35 13 0.12 (0.1) 46.2 1 0.15 (NA) 100.0 Negative Controls 25 0.05 (0.01) 0 20 0.04 (0.01) 0 Topical Contact 1 22 1.62 (0.1) 100.0 16 0.43 (0.1) 100.0 3 12 1.26 (0.6) 100.0 20 0.40 (0.1) 100.0 5 8 1.13 (0.5) 100.0 1 0.46 (NA) 100.0 8 13 1.26 (0.4) 100.0 2 0.64 (0.1) 100.0 12 16 1.23 (0.5) 100.0 8 0.45 (0.2) 100.0 13 3 0.66 (0.2) 100.0 3 0.41 (0.2) 100.0 15 3 0.30 (0.1) 100.0 0 --- --- 17 22 0.46 (0.3) 100.0 6 0.38 (0.3) 66.7 19 7 0.34 (0.3) 100.0 2 0.40 (0.1) 100.0 21 1 0.07 (NA) 100.0 1 0.04 (NA) 0.0 34 7 0.16 (0.1) 57.1 10 0.19 (0.2) 80.0 35 4 0.16 (0.2) 50.0 1 0.49 (0.3) 100.0 Negative Controls 20 0.05 (0.01) 0 20 0.04 (0.01) 0 - 258 -
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IMPACT OF HOST PLANT XYLEM FLUID ON
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0.18 600 Optical density 0.16 0.14
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OPTIMIZING MARKER-ASSISTED SELECTIO
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esistant and susceptible genotypes
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MAP BASED IDENTIFICATION AND POSITI
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esistant genotypes are in process a
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BREEDING PIERCE’S DISEASE RESISTA
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Progeny from crosses of field resis
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a = (1=low, 4= high); b = (1=green,
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v.8) for differences due to the pla
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SHARPSHOOTER FEEDING BEHAVIOR IN RE
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correlated with all other findings
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EFFECTS OF FEEDING SUBSTRATE ON RET
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REFERENCES Bextine, B. and T. Mille
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will also provide insights into the
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OBJECTIVES 1. Determine glassy-wing
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GWSS per 30 s sweep (seasonal avera
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ELISA for the presence of GWSS egg
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Project Leader: Thomas P. Freeman E
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Freeman, T. P., R. A. Leopold, D. R
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pathogen, when they move into viney
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A NOVEL IMMUNOLOGICAL APPROACH FOR
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1.6 GWSS Adults That Fed on Protein
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Hagler, J.R. & S.E. Naranjo. 2004.
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RESULTS: Oviposition Survey Wild gr
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Host specificity testing: No-choice
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currently employed morphological ch
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increase our present understanding
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BIOLOGY AND MORPHOMETRIC ANALYSIS O
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Table 1. Mean a developmental durat
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EFFECTS OF USING CONSTANT AND CYCLI
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REFERENCES Gautam, R. D. 1986. Effe
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PARASITISM OF THE GLASSY-WINGED SHA
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Table 1. Parasitism by G.ashmeadi o
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Project Leader: Robert F. Luck Dept
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A more interesting analysis using t
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MYCOPATHOGENS AND THEIR EXOTOXINS I
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POPULATION DYNAMICS AND INTERACTION
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No egg masses were recorded on olea
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EXPLORATION FOR FACULTATIVE ENDOSYM
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Although Baumannia and Wolbachia we
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EFFECTS OF SUBLETHAL DOSES OF IMIDA
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Table 2. Mortality and flight perfo
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A NOVEL METHOD TO INDUCE OVIPOSITIO
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REFERENCES Brodbeck, B. V., P. C. A
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Cohorts H. coagulata neonates were
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REFERENCES Blua, M. J. and D. J. W.
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Figure 2 shows the average numbers
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REPRODUCTIVE BIOLOGY AND PHYSIOLOGY
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REFERENCES Blua, M. J., Phillips, P
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RESULTS Some plant families had no
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Table 5. Winter, spring and summer
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16S rDNA is by far the most sequenc
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DNA MICROARRAY AND MUTATIONAL ANALY
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Inhibition of biofilm is BSA concen
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CULTURE-INDEPENDENT ANALYSIS OF END
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Borneman, J., M. Chrobak, G. Della
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P (CFU P OBJECTIVE 1. Determine the
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Purcell, AH and SR Saunders. 1999a.
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Figure 1: Southern blot of tolC::ap
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Project Leader: David Gilchrist Dep
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III. Production of transgenic plant
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RESULTS Construction of cDNA Librar
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CONCLUSIONS Genetic resistance and
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Mutagenesis of Xylella The EZ::TN T
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ISOLATION OF BACTERIOPHAGES SPECIFI
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Project Leader: Michele M. Igo Sect
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outer membrane fractions using two-
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1995; Purcell and Saunders, 1999).
Page 147 and 148: DEVELOPMENT OF SSR MARKERS FOR GENO
Page 149 and 150: Strain Name Host of Origin County o
Page 151 and 152: ROLE OF ATTACHMENT OF XYLELLA FASTI
Page 153 and 154: wild-type cells was not conclusive
Page 155 and 156: DETERMINATION OF GENES CONFERRING H
Page 157 and 158: S1 S2 S3 S4 Nb123456789bb123456789b
Page 159 and 160: MULTILOCUS SEQUENCE TYPING TO IDENT
Page 161 and 162: GENOME-WIDE IDENTIFICATION OF RAPID
Page 163 and 164: Americas and since most of the plan
Page 165 and 166: EFFECTS OF CHEMICAL MILIEU ON ATTAC
Page 167 and 168: CONCLUSIONS Our overall objective i
Page 169 and 170: RESULTS Objective 1. We conducted t
Page 171 and 172: Redak, R. A., Purcell, A. H., Lopes
Page 173 and 174: In parallel, an “activating trans
Page 175 and 176: PATTERNS OF XYLELLA FASTIDIOSA INFE
Page 177 and 178: % of vessels 100 80 60 40 20 Mugwor
Page 179 and 180: DOCUMENTATION AND CHARACTERIZATION
Page 181 and 182: Magnolia002 showed more identity (9
Page 183 and 184: PLASMID ADDICTION AS A NOVEL APPROA
Page 185 and 186: GENETIC VARIABILITY OF XYLELLA FAST
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Page 189 and 190: probing activities). In preliminary
Page 191 and 192: the slow release valve was opened a
Page 193 and 194: 12. Hopkins DL (1977) Diseases caus
Page 195 and 196: The almost complete absence of info
Page 197: QUANTIFYING LANDSCAPE-SCALE MOVEMEN
Page 201 and 202: EPIDEMIOLOGICAL ASSESSMENTS OF PIER
Page 203 and 204: multiply to relatively high (easily
Page 205 and 206: SPATIAL DATABASE CREATION AND MAINT
Page 207 and 208: Project Leader: Thomas M. Perring D
Page 209 and 210: Figure 2. Individual leaves from a
Page 211 and 212: The state variables, process functi
Page 213 and 214: DEVELOPMENT OF A FIELD SAMPLING PLA
Page 215 and 216: Figure 1. Three main dispersion pat
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Page 221 and 222: OBJECTIVES 1. Track the movement of
Page 223 and 224: REFERENCES 1. Beard, C.B., Cordon-R
Page 225 and 226: cases, positive phloem samples were
Page 227 and 228: EXPLOITING XYLELLA FASTIDIOSA PROTE
Page 229 and 230: Figure 2. Polymer disk accumulation
Page 231 and 232: CHARACTERIZATION OF NEONICOTINOIDS
Page 233 and 234: EVALUATION OF RESISTANCE POTENTIAL
Page 235 and 236: Project Leader: Doug Cook Dept. of
Page 237 and 238: Based on the in silico analysis, de
Page 239 and 240: CONTROL OF PIERCE’S DISEASE THROU
Page 241 and 242: Figure 1. Oleander ‘White’ afte
Page 243 and 244: RESULTS During the reporting period
Page 245 and 246: DEVELOPMENT OF AN ARTIFICIAL DIET A
Page 247 and 248: DESIGN OF CHIMERIC ANTI-MICROBIAL P
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Neutrophil elastase Cecropin B Chim
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and WTXb is very low (0.00-0.40%) a
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100 100 0.056 North America 100 0.0
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GENETIC DIFFERENTIATION AMONG GEOGR
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Table 1. Single-populations descrip
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MOLECULAR DISTINCTION BETWEEN POPUL
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These novel observations strongly s
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SEQUENCE DIVERGENCE IN TWO MITOCHON
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Table 3. Pairwise sequence distance
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DEVELOPMENT OF MOLECULAR DIAGNOSTIC
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A. B. Relative Density of SCAR Band
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THE ALIMENTARY TRACK OF GLASSY-WING
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We have dissected and identified al
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REALIZED LIFETIME PARASITISM AND TH
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REPRODUCTIVE AND DEVELOPMENTAL BIOL
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Mean adult longevity (days) 25 20 1
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the proposed exploratory work; thes
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SEARCHING FOR AND COLLECTING EGG PA
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REFERENCES Hoddle, M. S. and S. V.
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some Gram(+) bacteria, but are inac
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7. Hultmark, D., et al., Insect Imm
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Isolation of Fungal Pathogens Soil
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IDENTIFICATION OF MECHANISMS MEDIAT
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concentrations of 1.5 x 10 7 bacter
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anchor it in the outer membrane (sh
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SYMBIOTIC CONTROL OF PIERCE’S DIS
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MANAGEMENT OF PIERCE’S DISEASE OF
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pfF mutants are taken up by insects
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Simpson, A. J. G., F. C. Reinach, P
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Al-Wahaibi, A.K., Owen, and J. G. M
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patterns differed among the lines,
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Punja, Z.K. 2001. Genetic engineeri
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mind, we conducted an additional st
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REFERENCES Toscano, N., Byrne, F.,
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RESULTS AND CONCLUSIONS The program
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COMPATIBILITY OF INSECTICIDES WITH
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More tests are in progress to addre
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